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Microtubule Staining

Molecular light posts for cellular traffic routes
   



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Microtubules are tracks of cellular transport, permitting the well-ordered distribution of organelles over long distances. Assembly and disassembly of microtubule fibers are tightly regulated events that depend on the binding of GTP to tubulin dimers. To observe microtubule structures and transportation phenomena directly, high-resolution fluorescence microscopy techniques, e.g. TIRF, are the methods of choice. Towards this end, dye-labeled 2'/3'-O-(2-aminoethyl-carbamoyl) (= EDA)-GTP-derivatives (Scheme 1A) have been identified as ideal fluorescent GTP surrogates[1]. When supplemented to growing microtubules (Scheme 1B), Dye-EDA-GTPs allow

  • Highly specific and scalable staining
  • Flexibility in the choice of fluorophores
  • Labeling with little influence on motor protein kinetics

Scheme 1: A) Chemical Structure of Dye-EDA-GTP conjugates. B) Binding of Dye-EDA-GTP to the β-subunit of αβ-tubulin dimers induces polymerization to yield dye-labeled microtubules.

Dye-EDA-GTP conjugates for microtubule staining offered by Jena Bioscience:

Dye

λexem [nm]

Cat. No.

Amount

Price (€)

5/6-TAMRA

545/575

NU-820-TAM

160 µl (1 mM)

865,00

Cy3

550/570

NU-820-CY3

160 µl (1 mM)

865,00

Cy5

649/670

NU-820-CY5

160 µl (1 mM)

865,00

Fields of application:

Our tailored EDA-GTPs have been successfully applied in

  • Fluorescence/super-resolution microscopy on microtubules[1]
  • Photobleaching experiments[1]
  • Microtubule gliding assays[1-4]
  • The determination of microtubule persistence lengths[3,4]


Selected References:
[1] Anderson et al. (2011) A fluorescent GTP analog as a specific, high-precision label of microtubules. Biotechniques 51 (1):43.
[2] Fujimura et al. (2017) Dissection of the angle of single fluorophore attached to the nucleotide in corkscrewing microtubules. Biochem. Biophys. Res. Commun. 485 (3):614.
[3] Martin (2013) Measuring microtubule persistence length using a microtubule gliding assay. Methods Cell Biol. 115:13.
[4] Martin et al. (2012) Flexural rigidity measurements of biopolymers using gliding assays. J. Vis. Exp. 69:50117.

   
   
   
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